Synthesis of Sorafenib-Ruthenium Complexes, Investigation of Biological Activities and Applications in Drug Delivery Systems as an Anticancer Agent.

Sorafenib, a multiple kinase inhibitor, is widely used as a first-line treatment for hepatocellular carcinoma. However, there is a need for more effective alternatives when sorafenib proves insufficient. In this study, we aimed to design a structure that surpasses sorafenib's efficacy, leading us to synthesize sorafenib-ruthenium complexes for the first time and investigate their properties. Our results indicate that the sorafenib-ruthenium complexes exhibit superior epidermal growth factor receptor (EGFR) inhibition compared to sorafenib alone. Interestingly, among these complexes, Ru3S demonstrated high activity against various cancer cell lines including sorafenib-resistant HepG2 cells while exhibiting significantly lower cytotoxicity than sorafenib in healthy cell lines. Further evaluation of cell cycle, cell apoptosis, and antiangiogenic effects, molecular docking, and molecular dynamics studies revealed that Ru3S holds great potential as a drug candidate. Additionally, when free Ru3S was encapsulated into polymeric micelles M1, enhanced cytotoxicity on HepG2 cells was observed. Collectively, these findings position Ru3S as a promising candidate for EGFR inhibition and warrant further exploration for drug development purposes.

GLUT-Targeting Phototherapeutic Nanoparticles for Synergistic Triple Combination Cancer Therapy.

The combination of multimodal therapies into one nanocarrier system is promising for its potential to enhance treatment performance by overcoming the efficacy problems encountered in conventional monomodal therapy. In this study, targeted and multimodal therapeutic hybrid nanocarriers are fabricated for breast cancer treatments. In this context, the synthesized gold nanorods (AuNRd), photothermal therapy (PTT) agent, are coated with doxorubicin (DOX) conjugated, targeted, and biocompatible tetrablock glycopeptide (P(DMAEMA-b-HMBAMA-b-FrucMA)-b-P(Lys)/DOX, P-DOX) polymer. Here, fructose-based (Fruc) glycopeptide polymer enhances cellular uptake into breast cancer through GLUT5. A photosensitizer molecule, indocyanine green (ICG), was loaded into the particles to provide photodynamic therapy (PDT) upon NIR light at 808 nm. In the final step of the fabrication, the polymer-coated nanoparticles are integrated with antisense ISIS5132 oligonucleotides to prevent apoptotic resistance of cells against drug molecules. The biocompatibility and therapeutic efficacy of the nanoparticles are evaluated on both human normal skin fibroblast cell (CCD-1079Sk) and human breast cancer cell (MCF7) lines. These multimodal therapeutic AuNRd@P-DOX/ICG/ISIS5132 nanoparticles demonstrate an efficient triple synergistic effect of chemo-/PTT/PDT, which is desired for breast cancer treatment. We believe that this promising multimodal therapeutic nanoparticle system can promote the further clinical application in the treatment of breast cancer and can also be adapted to other types of cancer.

Synthesis and Comprehensive in Vivo Activity Profiling of Olean-12-en-28-ol, 3ß-Pentacosanoate in Experimental Autoimmune Encephalomyelitis: A Natural Remyelinating and Anti-Inflammatory Agent.

Multiple sclerosis (MS) treatment has received much attention, yet there is still no certain cure. We herein investigate the therapeutic effect of olean-12-en-28-ol, 3ß-pentacosanoate (OPCA) on a preclinical model of MS. First, OPCA was synthesized semisynthetically and characterized. Then, the mice with MOG35-55-induced experimental autoimmune/allergic encephalomyelitis (EAE) were given OPCA along with a reference drug (FTY720). Biochemical, cellular, and molecular analyses were performed in serum and brain tissues to measure anti-inflammatory and neuroprotective responses. OPCA treatment protected EAE-induced changes in mouse brains maintaining blood-brain barrier integrity and preventing inflammation. Moreover, the protein and mRNA levels of MS-related genes such as HLD-DR1, CCL5, TNF-α, IL6, and TGFB1 were significantly reduced in OPCA-treated mouse brains. Notably, the expression of genes, including PLP, MBP, and MAG, involved in the development and structure of myelin was significantly elevated in OPCA-treated EAE. Furthermore, therapeutic OPCA effects included a substantial reduction in pro-inflammatory cytokines in the serum of treated EAE animals. Lastly, following OPCA treatment, the promoter regions for most inflammatory regulators were hypermethylated. These data support that OPCA is a valuable and appealing candidate for human MS treatment since OPCA not only normalizes the pro- and anti-inflammatory immunological bias but also stimulates remyelination in EAE.

Dual-Drug Conjugated Glyco-Nanoassemblies for Tumor-Triggered Targeting and Synergistic Cancer Therapy.

Drug-conjugated nanoassemblies potentiate the efficiency of anticancer drugs through the advantages of high drug-loading capacity and passive/active targeting ability in cancer therapy. This study describes the synthesis of gemcitabine (Gem) and cisplatin (cisPt) dual-drug-functionalized glyco-nanoassemblies (GNs) for anticancer drug delivery systems. It also investigates the pH-triggered drug delivery of the conventional anticancer drug cisPt. A Gem-functionalized well-defined glycoblock copolymer backbone (P(iprFruMA-b-MAc)-Gem), which consists of fructose and methacrylic acid segments, was synthesized via a reversible addition-fragmentation chain transfer (RAFT) polymerization method. Following the hydrolysis of the protecting groups on the backbone copolymer, cisPt functionalization of P(FruMA-b-MAc)-Gem in aqueous media was carried out during the transformation of glycoblock polymers into self-assembled spherical glyco-nanoassemblies (GN3). Monodrug-functionalized glyco-nanoassemblies were also prepared either with Gem (GN1) or cisPt (GN2) to compare the synergetic effect of dual-drug conjugated glyco-nanoassemblies (GN3). The sizes of glyco-nanoassemblies GN1, GN2, and GN3 were found as 5.76 ± 0.64, 59.80 ± 0.13, and 53.80 ± 3.90 nm and dispersity (D) values as 0.476, 0.292, and 0.311 by dynamic light scattering (DLS) measurement, respectively. The in vitro studies revealed that the drug-free glyco-nanoassemblies are biocompatible at concentrations higher than 296 µg/mL. The drug-conjugated glyco-nanoassemblies (GN1 and GN2) exhibited in vitro cytotoxicity against human breast cancer cell lines of MDA-MB-231 comparable to free Gem and cisPt, illustrating an efficient drug release into the tumor environment. Additionally, GNs exhibited higher selectivity and preferential cellular internalization in MDA-MB-231 when compared to healthy cell lines of CCD-1079Sk. These dual-drug conjugated GNs can effectively enhance the killing of cancer cells and increase synergistic chemotherapy.

Sulforaphane, a Chemopreventive Compound Induces Necrotic Behavior and Inhibits S-phase of Cell Cycle in Human Kidney Cells in Vitro.

Sulforaphane (SFN) is an organosulfur product of found isothiocyanates in vegetables. The chemopreventive effects of SFN have revealed that there is a link between excessive consumption of SFN-rich vegetables and cancer formation without possible toxicological consequences. We aimed to evaluate the cellular outcome of SFN from a toxicological perspective, particularly for renal cells including clear cell adenocarcinoma (769-P) and human embryonic renal epithelial (293T) cells. The viability/cytotoxicity experiments were performed with methyl thiazole diphenyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. IC50-dependent, non-cytotoxic concentrations were used for the determination of cell cycle status and apoptosis by using flow cytometry and western blot. A certain concentration of SFN effectively altered apoptotic/necrotic behavior in 769-P compared to the control group 293T. Cell cycle status remained stable while showing a decreased proliferation profile for 769-P cells. The percentage of the S phase from the cell cycle in 293T cells significantly reduced without affecting proliferation status. The use of SFN as an alternative to traditional treatments might be considered for the battle against renal cell carcinoma but the current findings showed that caution should be applied particularly for renal cells. Our study will provide a basis for future in vivo studies to support traditional cancer therapies.

Increasing the biocompatibility of graphene-based hybrid nanostructures with glycopolymer.

Hybrid nanostructures decorated with glycopolymers are appropriate for biomedical applications. In this paper, the results are obtained from nanographene (NG) decorated with glycoblock copolymer to increase their potential in use in therapies and in examining lectin interactions. A pyren-1-ylmethyl 4-cyano-4-((phenylcarbonothioyl)thio)pentanoate (CPADB-py) chain transfer agent was used in the synthesis of methyl methacrylate glycoblock copolymers (P2 and P3) by reversible addition-fragmentation chain transfer (RAFT) polymerization to adhere the polymer to the nanographene surface. Hybrid nanographenes (NG-1 and NG-2) were obtained by the non-covalent interaction of deprotected P2 and P3 with different fructopyranose groups (3-O-methacryloyl-1,2:4,5-di-O-isopropylidene-ß-D-fructopyranose and 1-O-methacryloyl-2,3:4,5-di-O-isopropylidene-ß-D-fructopyranose) in their backbones. Images obtained from transmission electron microscopy (TEM) of NG-1 and NG-2 show that glycoblock copolymer coating was performed homogeneously. Moreover, thermal gravimetric analysis (TGA) also confirmed a glycoblock copolymer coating of NG-1 and NG-2 by weight loss of 41% and 31%, respectively. In the last step of the study, the binding ability of glycoblock copolymers (P2-hyd and P3-hyd) with concanavalin A (ConA) lectin was investigated by a turbidimetric assay. Promising results were obtained from P3-hyd for the ConA interactions. Hence, this study may open a new avenue in the design of new multifunctional glyconanomaterials that show favorable binding properties with lectins.

Evaluation of cytotoxic and antioxidant potential of Dittrichia viscosa (L.) Greuter used in traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Dittrichia viscosa (L.) Greuter ("Sari ot, Yapiskan andiz otu" in Turkish) is a medicinal plant that has been traditionally used in the Mediterranean area. This plant is used by the local population for the treatment of cancer. Investigation of their biological activities is therefore very important to be supported by scientific basis for traditional use. AIMS OF THE STUDY: In this study, it is aimed to assess the phytochemical composition, in vitro antioxidant, cytotoxic, and antiproliferative activities of the aqueous and ethanolic extracts obtained from the aerial parts (stems, leaves, flowers) of D. viscosa, collected from two sites in Turkey (Istanbul and Marmaris) against breast and prostate tumor cell lines. MATERIALS AND METHODS: Validated methods were used to evaluate the in vitro antioxidant capacity (DPPH, ABTS, CUPRAC), cytotoxicity (Cell Viability Assay), antiproliferative (Apoptosis assay), and phytochemical compositions. The nepetin (N), 3-O-methylquercetin (Q), and hispidulin (H) in the extracts of D. viscosa were quantified by HPLC and LC-HRMS. Furthermore, in order to control the standards of benefiting from the plant in a healthy way, the contents of some heavy metals were also assessed by ICP-OES in the plant and soil samples as well as the species soil's physical and chemical characteristics. RESULTS: We have found that heavy metal accumulation in the soil does not exceed the allowable limit value except for the nickel. The results showed that ethanol extraction is an efficient strategy to get NQH molecules with a higher content compared with other extraction techniques. However, using the same extraction method revealed that the amount of NQH molecules in the samples of two different regions were variable. The results suggested that all extracts had a high amount of total phenolic content (12.354-22.184 µg GAE/mg) and total flavonoid content (4.442-17.263 µg QE/g). In the antioxidant assay according to the DPPH method, the aqueous ethanol extracts (IC50; 21.00 µg/mL) showed stronger antioxidant activity than BHT. A significant reduction in cell viability was particularly observed in MDA-MB-231 cells, which were sensitive to ethanolic extracts in Istanbul (12-22%) and in Marmaris (14-15%), while PC3 cell lines were also more sensitive to extracts of the aqueous in Istanbul (16%) and the decoction in Marmaris (12%) after 72 h. Especially, it was observed that Marmaris and Istanbul samples induced the toxicity against PC3 cells. CONCLUSION: The study supports the medicinal use of D. viscosa as a potential anticancer against breast and prostate cancer cells in vitro and underlines the immense therapeutic potential of the plant.

Phthalocyanine-Conjugated Glyconanoparticles for Chemo-photodynamic Combination Therapy.

Combination cancer therapy based on multifunctional nanomaterials has attracted great attention. The present work focuses on the preparation of the glycopolymeric nanoparticle, which contains a photosensitizer (zinc(II)phthalocyanine, ZnPc) and an anticancer drug (Doxorubicin, Dox). First, a novel mono azide-functional ZnPc-N3 with seven hydrophilic ethylene oxide chains was synthesized. Next, ZnPc alone or together with Dox bearing glycopolymers was synthesized via the RAFT polymerization method and then self-assembled into glyconanoparticles (GNPs) with narrow particle size distribution. Then the evaluation of the biological activity of GNPs (GNPs-ZnPc and GNPs-ZnPc/Dox) for dual photodynamic therapy (PDT) and chemotherapy against human breast cancer cells was investigated. The constructed GNPs were identified via general characterization methods, including dynamic light scattering (DLS) and transmission electron microscopy (TEM). The prepared GNPs-ZnPc/Dox demonstrated remarkable photophysical and photochemical properties, involving good colloidal stability in biological conditions, pH-responsive drug release, and the capacity to generate singlet oxygen under light irradiation. The outer layer of nanoparticles covered by fructose sugar moieties achieves a targeted cancer therapy owing to GLUT5 (a well-known fructose transporter) overexpression toward breast cancer cells. In vitro experiments were then performed to evaluate the chemo/phototoxicity, cellular uptake, and anticancer efficacy of GNPs-ZnPc/Dox. In comparison with free Dox, human breast cancer cells treated with GNPs-ZnPc/Dox exhibited a higher cellular internalization via GLUT5 targeting. In particular, the GNPs-ZnPc/Dox nanoplatform revealed an excellent synergistic anticancer activity in comparison with free ZnPc-N3 and free Dox, representing a novel and promising chemo-photodynamic combination therapeutic methodology to improve therapeutic efficacy.

Glycopolymer decorated multiwalled carbon nanotubes for dual targeted breast cancer therapy.

Carbon-based nanomaterials (CNMs) have attracted great attention in biomedical applications such as cancer imaging and therapy. CNMs, which are currently used in a wide range of applications, suffer from drawbacks of toxicity and low biocompatibility. Either noncovalent or covalent functionalization of CNMs with hydrophilic and biocompatible polymers which help to block hydrophobic interactivity between CNMs and cells can greatly increase their biocompatibility by eliminating their probable toxicity towards living organisms. In this report, we present a comparison of both noncovalent and covalent functionalization approaches in order to introduce a biocompatible glycoblock copolymer onto multi-walled carbon nanotubes (CNTs) in order to enhance their potential in therapies. An anticancer drug (doxorubicin, Dox) was conjugated with two different end functionalized poly(1-O-methacryloyl-ß-d-fructopyranose-b-(2-methacryloxyethoxy))benzaldehyde glycoblock copolymers, which were synthesized via reversible addition-fragmentation chain transfer (RAFT) polymerization, by either noncovalent or covalent tethering. CNTs were coated separately with the synthesized drug-conjugated glycoblock copolymers and folic acid (FA) to obtain an efficient drug delivery platform for dual-targeting of glucose transporter protein (GLUT5) and folic acid receptors (FR) in breast cancer. A library of synthesized monomers, polymers and prepared glycoblock copolymer coated CNTs (hybrid-CNTs) using both approaches were comprehensively characterized by various techniques. Transmission electron microscopy measurements showed the homogeneous, smooth morphology of the prepared Dox-conjugated glycoblock copolymer coating of CNTs and confocal laser scanning microscopy images displayed successful cellular internalization of hybrid-CNTs in the MCF-7 and MDA-MB-231 human breast cancer cell lines. This research demonstrates the potential of hybrid-CNTs as a biocompatible drug delivery system as well as in vitro use of Dox-conjugated vehicles for dual receptor mediated breast cancer therapy.

Folic Acid-Conjugated pH and Redox-Sensitive Ellipsoidal Hybrid Magnetic Nanoparticles for Dual-Triggered Drug Release.

Stimuli-sensitive and multifunctional nanoparticles are highly desirable biomedical materials for triggered and targeted drug delivery applications. Here, we have designed pH- and redox-triggered magnetic lipid-polymer hybrid nanoparticles (MHNPs) with a core-shell structure. This design is composed of a silica-/mesoporous silica-coated ellipsoidal magnetic nanoparticle with multifunctionality: carrying the anticancer drug (doxorubicin, DOX), the cancer cell targeting ligand (folic acid-conjugated poly(ethylene glycol), FA-PEG) polymer, and being coated with a biocompatible pH-/redox-responsive (poly-l-histidine-poly(ethylene glycol)-lipoic acid; PLH-PEG-LA) polymer. The lipoic acid units of the PLH-PEG-LA shell of the FA-MHNPs were cross-linked using 1,4-dithiothreitol (DTT). When the FA-MHNPs-DOX were exposed to an endolysosomal pH of 5.5 and 10 mM glutathione (GSH), the particles exhibited a very efficient DOX release profile within 24 h. In addition, cytotoxicity, uptake, and apoptosis assays were performed against breast cancer cell lines. These results showed that FA-MHNPs-DOX promote an enhanced uptake and cell morbidity compared to the nontargeted MHNPs-DOX against tested cell lines. Moreover, the FA-MHNPs-DOX exhibited very effective cytotoxicity and also decreased the cell viability through apoptosis against breast cancer cell lines. In conclusion, it can be said that the pH and redox dual-responsive hybrid FA-MHNPs-DOX has a great potential for controlled drug release.

Glyconanoparticles for Targeted Tumor Therapy of Platinum Anticancer Drug.

An important requirement to decrease the side effects of chemotherapy drugs is to develop nanocarriers with precise biological functions. In this work, a set of glyconanoparticles was prepared via self-assembly of amphiphilic glycoblock copolymers for the targeted delivery of a hydrophobic chemotherapy drug. Well-defined glycoblock copolymers that consist of 1,1-di-tert-butyl 3-(2-(metyloyloxy)ethyl)-butane-1,1,3-tricarboxylate (MAETC) together with three different protected-sugar moieties (ß-d-glucopyranoside, ß-d-mannopyranoside, and ß-l-fucopyranoside) were synthesized by using reversible addition-fragmentation chain-transfer polymerization. Copolymers were deprotected and conjugated with the cis-dichlorodiammineplatinum(II) (cis-Pt) anticancer drug. Dynamic light scattering and transmission electron microscopy measurements revealed that cis-Pt-conjugated glyconanoparticles were sufficiently stable under physiological conditions and had diameters of approximately 100 nm with considerably narrow size distributions. They were intracellularly taken up by the breast cancer (MCF-7 and MDA-MB-231), prostate cancer (PC3), renal cancer (769-P), and Chinese hamster ovary cell lines. The PC3 and 769-P cell lines showed a high preference for the glycosylated nanoparticles. Glycoblock copolymers were found nontoxic but showed high cytotoxicity and increased efficacy after conjugation with the cis-Pt anticancer drug. Moreover, in vitro cytotoxicity assays in cancer cell lines demonstrate that cis-Pt-loaded glycopolymer-based nanoparticles have higher cytotoxicity than free cis-Pt. Overall, our results suggest that glyconanoparticles have a great potential to be used as an effective cis-Pt drug carrier for targeted cancer therapy.

Synthesis, biological activity and multiscale molecular modeling studies of bis-coumarins as selective carbonic anhydrase IX and XII inhibitors with effective cytotoxicity against hepatocellular carcinoma.

A series of novel bis-coumarin derivatives containing triazole moiety as a linker between the alkyl chains was synthesized and their inhibitory activity against the human carbonic anhydrase (hCA) isoforms I, II, IX and XII were evaluated. In addition, cytotoxic effects of the synthesized compounds on renal adenocarcinoma (769P), hepatocellular carcinoma (HepG2) and breast adeno carcinoma (MDA-MB-231) cell lines were examined. While the hCA I and II isoforms were inhibited in the micromolar range, the tumor-associated isoform hCA IX and XII were inhibited in the high nanomolar range. 4-methyl-7-((1-(12-((2-oxo-2H-chromen-7-yl)oxy)dodecyl)-1H-1,2,3-triazol-4-yl)methoxy)-2H-chromen-2-one (5p) showed the strongest inhibitory activity against hCA IX with the Ki of 144.6 nM and 4-methyl-7-((1-(10-((2-oxo-2H-chromen-7-yl)oxy)decyl)-1H-1,2,3-triazol-4-yl)methoxy)-2H-chromen-2-one (5n) exhibited the highest hCA XII inhibition with the Ki of 71.5 nM. In order to better understand the inhibitory profiles of studied molecules, multiscale molecular modelling approaches were applied. Low energy docking poses of studied molecules at the binding sites of targets have been predicted. In addition, electrostatic potential surfaces (ESP) for binding sites were also generated to understand interactions between proteins and active ligands.

Assessment of in vitro cytotoxic and genotoxic activities of some trimethoprim conjugates.

Trimethoprim, a commonly used antibacterial agent, is widely applied in the treatment of variety of infections in human. A few studies have demonstrated an extensive exposure of man to antibiotics, but there is still a lack of data for cytotoxic effects including nephrotoxicity, gastrointestinal toxicity, hematotoxicity, neurotoxicity and ototoxicity. The main purpose behind this study was to determine cytotoxic and genotoxic activities of trimethoprim (1), trimethoprim with maleic acid (2) and trimethoprim in conjugation with oxalic acid dihydrate (3). The cytotoxic effects of these three conjugates were elucidated by employing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoium bromide (MTT) assay using embryonic rat fibroblast-like cell line (F2408) and H-ras oncogene activated embryonic rat fibroblast-like cancer cell line (5RP7). Additionally, determination of genotoxic activity of these three compounds were studied by using cytokinesis blocked micronucleus assay (CBMN) in human lymphocytes. The results demonstrated that trimethoprim alone and its combination with other compounds are able to induce both cytotoxic and genotoxic damage on cultured cells (F2408, 5RP7, human lymphocytes).

Design, synthesis and docking study of novel coumarin ligands as potential selective acetylcholinesterase inhibitors.

New coumaryl-thiazole derivatives with the acetamide moiety as a linker between the alkyl chains and/or the heterocycle nucleus were synthesized and in vitro tested as acetylcholinesterase (AChE) inhibitors. 2-(diethylamino)-N-(4-(2-oxo-2H-chromen-3-yl)thiazol-2-yl)acetamide (6c, IC50 value of 43 nM) was the best AChE inhibitor with a selectivity index of 4151.16 over BuChE. Kinetic study of AChE inhibition revealed that 6c was a mixed-type inhibitor. Moreover, the result of H4IIE hepatoma cell toxicity assay for 6c showed negligible cell death. Molecular docking studies were also carried out to clarify the inhibition mode of the more active compounds. Best pose of compound 6c is positioned into the active site with the coumarin ring wedged between the residues of the CAS and catalytic triad of AChE. In addition, the coumarin ring is anchored into the gorge of the enzyme by H-bond with Tyr130.

Synthesis, anticholinesterase activity and molecular modeling study of novel carbamate-substituted thymol/carvacrol derivatives.

New thymol and carvacrol derivatives with the carbamate moiety were synthesized and their inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) were evaluated. 5-isopropyl-2-methylphenyl(3-fluorophenyl)carbamate (29) was found to be the most potent AChE inhibitor with IC50 values of 2.22µM, and 5-isopropyl-2-methylphenyl (4-fluorophenyl)carbamate (30) exhibited the strongest inhibition against BuChE with IC50 value of 0.02µM. Additionally, the result of H4IIE hepatoma cell toxicity assay for compounds 18, 20, 29, 30 and 35 showed negligible cell death at 0.07-10µM. Moreover in order to better understand the inhibitory profiles of these molecules, molecular modeling studies were applied. Binding poses of studied compounds at the binding pockets of AChE and BuChE targets were determined. Predicted binding energies of these compounds as well as structural and dynamical profiles of molecules at the target sites were estimated using induced fit docking (IFD) algorithms and post-processing molecular dynamics (MD) simulations methods (i.e., Molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) approaches).

Cytotoxic, Apoptotic and Genotoxic Effects of Lipid-Based and Polymeric Nano Micelles, an In Vitro Evaluation.

Self-assembly systems (SAS) mainly consist of micelles, and liposomes are the classes of Nano Drug Delivery Systems with superior properties compared to traditional therapeutics in targeting cancer tumors. All commercially available nano-formulations of chemotherapeutics currently consist of SAS. According to our knowledge, a specific toxicity comparison based on material differences has not yet been performed. The purpose of this study was to evaluate and compare the toxicity of two SAS consisting of Sterically Stabilized Micelles (SSM) made of a lipid-based amphiphilic distearoyl-sn-glycero-phosphatidylethanolamine-polyethylene glycol (PEG)-2000 and a polymeric micelle (PM) consisting of Y-shape amphiphilic block copolymer, synthesized using poly ε-caprolactone and PEG. The mechanism of cytotoxicity and genotoxicity of micelles on L-929 healthy mouse fibroblast cells was assessed using Sulforhodamine-B, WST-1, Acridine Orange/Ethidium Bromide and alkaline single-cell gel electrophoresis assays. Results showed that SSM in conc. of 40 mg/mL shows very low cytotoxicity at the end of 24, 48 and 72 h. The DNA damage caused by SSM was much lower than PM while the latter one showed significant toxicity by causing apoptosis with the ED50 value of 3 mg/mL. While the DNA damage caused by SSM was ignorable, some DNA chain breaks were detected on cells treated with PM.

Albumin-polymer conjugate nanoparticles and their interactions with prostate cancer cells in 2D and 3D culture: comparison between PMMA and PCL.

Using proteins as the hydrophilic moiety can dramatically improve the biodegradability and biocompatibility of self-assembled amphiphilic nanoparticles in the field of nanomedicine. In this study, we fabricated and evaluated curcumin loaded albumin-polycaprolactone nanoparticles as a novel drug delivery system for prostate carcinoma therapeutics and compared their performance to poly(methyl methacrylate) (PMMA), a non-degradable and amorphous polymer. The maleimide functionalized poly(ε-caprolactone) (PCL) was obtain using ring opening polymerization (ROP) of ε-caprolactone where N-(2-hydroxyethyl)maleimide was used as an initiator. The resorbable albumin-polymer conjugate was prepared by conjugating the hydrophobic maleimide-terminated PCL to the hydrophilic bovine serum albumin (BSA) via a simple Michael addition reaction. PMMA was conjugated in a similar manner. The amphiphilic BSA-polymer conjugates can self-assemble into nanoparticles, displaying well-defined structure, prolonged storage stability, and excellent biocompatibility. The BSA nanoparticles, with encapsulated curcumin, exhibited highly enhanced antitumor activity compared to free curcumin. Furthermore, the high efficacy of the curcumin loaded nanoparticles was verified by effectively inhibiting the growth of three-dimensional LNCaP multicellular tumour spheroids. The cytotoxicity was attributed to the efficient cellular uptake of the nanoparticles through caveolic endocytosis. The direct comparison between PCL and the PMMA revealed that drug loading and release as well as cytotoxicity is not significantly affected by the nature of the polymer. However, it seems that nanoparticles based on PMMA penetrate quicker into LNCaP multicellular tumour spheroids thanks to the increased stability. The faster penetration was found to reduce the toxicity of the nanoparticles as evidenced by the lower number of dead cells. In contrast, the fully degradable PCL-based nanoparticles were more efficient in delivering the drug, thus limiting the growth of LNCaP multicellular tumour spheroids.

Carbohydrate-Specific Uptake of Fucosylated Polymeric Micelles by Different Cancer Cell Lines.

Inspired by upregulated levels of fucosylated proteins on the surfaces of multiple types of cancer cells, micelles carrying ß-l-fucose and ß-d-glucose were prepared. A range of block copolymers were synthesized by reacting a mixture of 2-azidoethyl ß-l-fucopyranoside (FucEtN3) and 2-azideoethyl ß-d-glucopyranoside (GlcEtN3) with poly(propargyl methacrylate)-block-poly(n-butyl acrylate) (PPMA-b-PBA) using copper-catalyzed azide-alkyne cycloaddition (CuAAC). Five block copolymers were obtained ranging from 100 mol % fucose to 100% glucose functionalization. The resulting micelles had hydrodynamic diameters of around 30 nm. In this work, we show that fucosylated micelles reveal an increased uptake by pancreatic, lung, and ovarian carcinoma cell lines, whereas the uptake by the healthy cell lines (CHO) is negligible. This finding suggests that these micelles can be used for targeted drug delivery toward cancer cells.

Dual-Responsive pH and Temperature Sensitive Nanoparticles Based on Methacrylic Acid and Di(ethylene glycol) Methyl Ether Methacrylate for the Triggered Release of Drugs.

A series of thermo-and pH-responsive poly(methyl methacrylate)-block-poly[methacrylic acid-co-di(ethylene glycol) methyl ether methacrylate] PMMA-b-P[MAA-co-DEGMA] block copolymers were synthesized by RAFT polymerization and self-assembled into micelles. The molar ratio of MAA was altered from 0-12% in order to modulate the lower critical solution temperature (LCST) of PDEGMA. The release of the drug albendazole from the micelle was strongly dependent on the temperature and the LCST value of the polymer. Systems below the LCST released the drug slowly while increasing the temperature above the LCST or decreasing the pH value to 5 resulted in the burst-like release of the drug. ABZ delivered in this pH-responsive drug carrier had a higher toxicity than the free drug or the drug delivered in a non-responsive drug carrier.

Polymer-albumin conjugate for the facilitated delivery of macromolecular platinum drugs.

The delivery of macromolecular platinum drugs into cancerous cells is enhanced by conjugating the polymer to albumin. The monomers N-(2-hydroxypropyl)methacrylamide (HPMA) and Boc protected 1,3-diaminopropan-2-yl acrylate (Ac-DAP-Boc) are copolymerized in the presence of a furan protected maleimide functionalized reversible addition-fragmentation chain transfer (RAFT) agent. The resulting polymer with a composition of P(HPMA14 -co-(Ac-DAP-Boc)9 ) and a molecular weight of Mn = 7600 g mol(-1) (D = 1.24) is used as a macromolecular ligand for the conjugation to the platinum drug. Thermogravimetric analysis reveals full conjugation. After deprotection of the maleimide functionality of the polymer, the reactive polymer is conjugated to albumin using the Cys34 functionality. The conjugation is monitored using size exclusion chromatography, MALDI-TOF (matrix assisted laser desorption ionization time-of-flight), and SDS Page (sodium dodecyl sulphate polyacrylamide gel electrophoresis). The polymer-albumin conjugates self-assemble in water into nanoparticles of sizes of around 80 nm thanks to the hydrophobic nature of the platinum drugs. The albumin coated nanoparticles are readily taken up by ovarian cancer cell lines and they show superior toxicity compared to a control sample without protein coating.